Journal: Scientific Reports

Article Title: Antagomir-mediated suppression of microRNA-134 reduces kainic acid-induced seizures in immature mice

doi: 10.1038/s41598-020-79350-7

Figure Lengend Snippet: Ant-134 has protective effects in the hippocampus and de-represses cortical DCX. ( a ) Representative FJB-stained images show neuronal death in the hippocampus 24 h after SE ipsi- and contralateral to pre-treatment with 0.1 nmol Ant-134 or scramble control (Scr). ( b ) The number of FJB-stained cells was reduced in mice pre-treated with Ant-134 (N = 6) compared with scramble control (N = 4). ( c ) Partial knockdown of miR-134 levels (54.8%) was retained 24 h after SE in the hippocampus of mice that were pre-treated with 0.1 nmol Ant-134 (N = 6) or Scr (N = 6). ( d ) Representative immunohistochemistry staining with Iba1 and GFAP in the CA3 of the hippocampus after SE. ( e ) The number of activated microglia was decreased in mice pre-treated with Ant-134 (N = 3) compared with scramble control (N = 3). ( f ) There was no difference in the number of astrocytes in mice pre-treated with Ant-134 (N = 3) and scrambled control (N = 3). ( g ) Western blot showing DCX expression in hippocampus and cortex at 24 h after KA-induced seizures of mice pre-treated with Scr/Ant-134 (0.1 nmol). Full-length blot can be found in Supplementary Fig. a. ( h ) Densitometry analysis of DCX in the hippocampus. GAPDH was used as the loading control, (N = 4/group). ( i ) Densitometry of DCX in the cortex. GAPDH was used as the loading control (N = 4/group).

Article Snippet: Following quenching and blocking, samples were incubated in primary antibodies GFAP (1:500, Biotechne 29415) and Iba1 (1:500, Wako Chemicals 19,741) overnight at 4 °C.

Techniques: Staining, Immunohistochemistry, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: Samples used in this study.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: In situ hybridisation for HtrA1 mRNA in keloid and normal skin. Sections from active keloid lesions ( a ) or unaffected region ( b ) (patient No. 18 in ) were hybridised with a probe specific to HtrA1 mRNA. Positive signals are visualised in blue. Scale bar = 50 µm.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: In Situ, Hybridization

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: An abdominal keloid after laparoscopic surgery. The activity of keloid was in order a, b and c. Higher activity in regions of the lesion were associated with increased cell proliferation and greater up-regulation of HtrA1. Scale bar = 500 µm.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: Immunohistochemical staining of HtrA1 protein in keloid ( a ) and normal skin tissue ( b ). Sections from active keloid lesions ( a ) or normal skin ( b ). ( a ) displays the results from patient No. keloid-3 in , and ( b ) displays the results from patient No. normal skin-1 in . Positive signals are visualised in brown. Scale bar = 50 µm.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: Western blot analysis of HtrA1 in keloid lesions and normal skin tissues. Soluble protein extract (8 µg/lane) was analysed using specific antibodies against HtrA1 or glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Keloid and normal skin samples from four different patients were analysed.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: Proportion of fibroblasts expressing HtrA1 protein in keloid lesions and normal skin. The number of fibroblasts with positive signals was counted after immunohistochemical staining of HtrA1 using samples from 17 keloidand 4 unrelated patients. Ten high-power (×400) fields were selected at random from a section and numbers of total and stained fibroblasts were counted. Patient information is described with proportion of HtrA1-positive cells in .

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: Proliferation rates of keloid fibroblasts and normal fibroblasts transfected with HtrA1 siRNA or control siRNA. Proliferation curves of keloid fibroblasts obtained from keloid sample No. 26 as shown in ( a ), ( n = 3) and normal fibroblasts from sample No. 8 ( d ) transfected with HtrA1 siRNA (knockdown) or control siRNA (control). The efficiency of HtrA1 knockdown in keloid fibroblasts was determined using western blot analysis ( b ) and quantitative PCR ( c ), ( n = 3). The efficiency of HtrA1 knockdown in normal fibroblasts was similarly determined using quantitative PCR ( e ), n = 3. Cell proliferation was analysed using a colorimetric assay with a water-soluble tetrazolium salt as the substrate. Error bars represent standard deviations ( n = 3). * p < 0.001.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: Transfection, Control, Knockdown, Western Blot, Real-time Polymerase Chain Reaction, Colorimetric Assay

Journal: International Journal of Molecular Sciences

Article Title: HtrA1 Is Specifically Up-Regulated in Active Keloid Lesions and Stimulates Keloid Development

doi: 10.3390/ijms19051275

Figure Lengend Snippet: Proliferation rates of keloid fibroblasts and normal fibroblasts incubated with or without recombinant HtrA1. Proliferation curves of keloid fibroblasts obtained from sample No. 29 and normal fibroblasts from sample No. 8 as shown in , incubated with (rHtrA1) or without (control) recombinant HtrA1. n = 3, * p < 0.01.

Article Snippet: The membranes were blocked with 5% Block Ace (DS Pharma Biomedical, Osaka, Japan) in PBS containing 0.05% Tween 20 prior to incubation with anti HtrA1 antibody (1:500, R&D Systems).

Techniques: Incubation, Recombinant, Control